Developing preclinical human iPSC-based HTS assays to identify therapeutic agents for biliary atresia

开发基于人类 iPSC 的临床前 HTS 检测，以确定胆道闭锁的治疗药物

• 批准号: 10206130
• 负责人: Yoon Young Jang
• 金额: $ 36.84万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10206130
• 关键词: Adverse effects; Automation; Basic Science; Biliary; Biliary Atresia; Biological Assay; Birth; Blood specimen; COL1A1 gene; Cell Line; Cell model; Cell physiology; Cells; Characteristics; Child; Childhood; Cirrhosis; Clinical; Collaborations; Collagen; Development; Disease; Disease model; Dose; Ensure; FDA approved; Fibrosis; Future; Hepatobiliary; Hepatotoxicity; Human; In Vitro; Infant; Inherited; Instruction; Libraries; Life; Liver; Liver Fibrosis; Liver diseases; Medical; Methods; Modeling; Morbidity - disease rate; Mus; Operative Surgical Procedures; Patients; Performance; Pharmaceutical Preparations; Phenotype; Process; Regenerative Medicine; Reporter; Reproducibility; Research; Resources; Safety; Source; System; Therapeutic; Therapeutic Agents; Tissues; Translating; Triage; alpha 1-Antitrypsin; alpha 1-Antitrypsin Deficiency; base; design; disease phenotype; falls; follow-up; high throughput screening; human disease; induced pluripotent stem cell; induced pluripotent stem cell technology; liver transplantation; miniaturize; mortality; mutant; novel; novel therapeutics; palliative; pre-clinical; prevent; prominin; response; success; therapeutic development

Project Summary The objective of the research is to develop patient iPSC-based HTS assays that can facilitate therapeutic discovery for treating biliary atresia (BA) fibrosis. BA is the most common cause of pediatric end-stage liver disease in the U.S. and is inevitably fatal within the first two years of life if untreated. Notably, BA is the most rapidly fibrosing liver disease in humans and is associated with significant morbidity and mortality in children. Compared to other liver diseases that gradually progress into cirrhosis over decades, BA infants characteristically develop fibrosis/cirrhosis within weeks to months after birth. Although there is a palliative surgical procedure, there is no known treatment to halt the progressive fibrosis; most infants born with the disease will need liver transplantation in order to survive. A main challenge in developing effective anti-fibrotic drugs has been the lack of a model of the human disease. The human induced pluripotent stem cell (iPSC) technology provides an alternative for generating functional, renewable and relevant cell sources for disease modeling using patient tissues. Based on our expertise on in vitro disease modeling, we have recently succeeded in developing BA patient- specific iPSCs and have demonstrated that these cells produce significantly more collagen and other fibrosis markers along with deficiency in biliary differentiation (key disease features of BA), compared to the iPSCs of healthy children. This new line of research on BA patient iPSCs makes it feasible to evaluate both efficacy and safety of potential drugs in a more human-relevant setting. Thus we believe this human cellular model of BA can serve as an ideal system to identify effective anti-fibrotic compounds in treating liver fibrosis in BA. In the current study, we propose to: 1) Develop a novel high throughput assay to assess anti-fibrotic effects of compounds on BA fibrosis using COL1A1 reporter BA-iPSC lines. We will determine the conditions for miniaturizing the assay and for robust assay automation. 2) Perform pilot screens to validate and optimize the assay using a clinical drug library in order to ensure automation reliability and assay reproducibility. 3) Develop independent secondary assays to prioritize hit selection by further verifying the anti-fibrotic effects of the hits and evaluating their protective/adverse effects on hepatobiliary tissues derived from patient iPSCs. At the conclusion of this study, we will have developed a robust, patient cell-based HTS assay capable of identifying new disease targets and leads for developing novel therapies for BA patients. Moreover, success of this project will be a step forward in translating basic iPSC discoveries to therapeutic applications, helping to fulfill their promise in developing regenerative medicine.

项目摘要 该研究的目的是开发基于IPSC的患者的HTS分析，以促进 治疗胆道闭锁（BA）纤维化的治疗发现。 BA是小儿最常见的原因 美国末期肝病，如果未经治疗，在生命的头两年内不可避免地致命。 值得注意的是，BA是人类中最快的纤维化肝病，与显着有关 儿童的发病率和死亡率。与其他肝病逐渐发展为 肝硬化数十年来，BA婴儿在数周到几个月内特征性地发展纤维化/肝硬化 出生后。尽管有姑息手术程序，但尚无已知治疗方法可以停止 进行性纤维化；大多数患有这种疾病的婴儿都需要肝移植才能 存活。开发有效抗纤维化药物的主要挑战是缺乏模型 人类疾病。 人类诱导的多能干细胞（IPSC）技术提供了生成的替代方法 使用患者组织进行疾病建模的功能，可再生和相关的细胞来源。基于 我们在体外疾病建模方面的专业知识，我们最近成功地发展了BA患者 - 特定的IPSC，并证明这些细胞产生的胶原蛋白和其他明显更多 与 健康孩子的IPSC。这项对BA患者IPSCS的新研究使其可行 在更相关的环境中评估潜在药物的功效和安全性。因此，我们相信 这种BA的人类细胞模型可以用作识别有效抗纤维化的理想系统 Ba中治疗肝纤维化的化合物。 在当前的研究中，我们建议：1）开发一种新型的高通量测定法以评估抗纤维化 使用COL1A1报告基因BA-IPSC线对化合物对BA纤维化的影响。我们将确定 小型化测定和可靠测定自动化的条件。 2）执行试点屏幕 使用临床药物库验证和优化测定法，以确保自动化可靠性和 测定可重复性。 3）开发独立的次要测定，以进一步确定命中选择 验证命中的抗纤维化作用，并评估其对其保护/不利影响 源自患者IPSC的肝胆组织。在这项研究的结论中，我们将 开发了一种强大的，基于患者细胞的HTS测定法，能够鉴定新的疾病靶标和铅 用于为BA患者开发新型疗法。此外，该项目的成功将是向前迈出的一步 在将基本的IPSC发现转化为治疗应用程序时，有助于实现他们的诺言 开发再生医学。